High spatio‐temporal resolution measurement of A1R and A2AR interactions combined with Iem‐spFRET and E‐FRET methods. Issue 11 (10th August 2021)

• Record Type: Journal Article
• Authors: Lin, Yating; Wang, Haoyu; Xu, Jianshu; Huang, Yiming; Gong, Wei; Wang, Qiwen; Huang, Zufang; Xie, Shusen; Lin, Juqiang
• Is Part Of: Journal of biophotonics. Volume 14:Issue 11(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-08-10
• Subjects: adenosine receptor -- cell membrane -- fluorescence resonance energy transfer (FRET) -- fluorescence spectrum; Photonics -- Periodicals; Optical materials -- Periodicals; Optics -- Periodicals; Medical instruments and apparatus -- Periodicals; 621.3605
• DOI: 10.1002/jbio.202100172
• Languages: English
• ISSNs: 1864-063X

Abstract

Abstract: A1 R‐A2A R heterodimers regulate striatal glutamatergic neurotransmission. However, few researches about kinetics have been reported. Here, we combined Iem‐spFRET and E‐FRET to investigate the kinetics of A1 R and A2A R interaction. Iem‐spFRET obtains the energy transfer efficiency of the whole cell. E‐FRET gets energy transfer efficiency with high spatial resolution, whereas, it was prone to biases because background was easily selected due to manual operation. To study the interaction with high spatio‐temporal resolution, Iem‐spFRET was used to correct the deviation of E‐FRET. In this paper, A1 R and A2A R interaction was monitored, and the changes of FRET efficiency of the whole or/and partial cell membrane were described. The results showed that activation of A1 R or A2A R leads to rapid aggregation, inhibition of A1 R or A2A R leads to slow segregation, and the interaction is reversible. These results demonstrated that combination of Iem‐spFRET and E‐FRET could measure A1 R and A2A R interaction with high spatio‐temporal resolution. Abstract : Iem‐spFRET and E‐FRET are popular quantitative methods for molecular interactions in living cells. Iem‐spFRET is a robust and independent emission‐spectral unmixing FRET methodology and circumvents the rigorous restriction of keeping the same imaging conditions for all FRET experiments, which tremendously improves the success rate of FRET experiment in single live cells. E‐FRET visualizes spatio‐temporal dynamics of signal molecules in living cells and can obtain efficiency of any region of interest (ROI). In this study, Iem‐spFRET combined with E‐FRET dynamically monitored CCPA (A1 R agonist)‐induced interaction between A1 R and A2A R, which obtain high spatio‐temporal resolution of the FRET efficiency value.